颗粒蛋白前体(PGRN)敲除小鼠的巨噬细胞抑制乳腺癌细胞侵袭和迁移的分子机制

Molecular mechanism of macrophages derived from PGRN gene knockout mice inhibit invasion and migration of breast cancer cells

目的探究颗粒蛋白前体基因敲除(PGRN-7-)小鼠的巨噬细胞与乳腺癌细胞侵袭和迁移的关系及机制。方法选择野生型和PGRN^-/-小鼠巨噬细胞条件培养基培养乳腺癌细胞,采用Transwell™实验、划痕实验检测癌细胞的侵袭、迁移能力,Western blot法检测上皮钙黏蛋白(E-cadherin)、神经钙黏蛋白(N-cadherin)的表达。通过细胞因子芯片、实时定量PCR和ELISA检测野生型和PGRN^-/-小鼠巨噬细胞分泌的细胞因子差异;用筛选出的差异表达细胞因子白细胞介素6(IL-6)处理乳腺癌细胞,采用上述方法检测对癌细胞侵袭迁移能力的影响并采用Western blot法检测核因子kB(NF-kB)信号通路和Janus激酶/信号转导子和转录激活子3(JAK/STAT3)信号通路在其中的作用。结果PGRN%小鼠的巨噬细胞NF-kB信号通路被阻断,减少IL-6分泌,抑制乳腺癌细胞侵袭和迁移;IL-6激活JAK/STAT3信号通路促进乳腺癌细胞侵袭迁移。结论PGRN^-/-小鼠的巨噬细胞NF-kB信号通路被阻断,下调ILE表达,进而抑制JAK/STA13信号通路的激活,抑制乳腺癌细胞的侵袭和迁移。

Objective To explore the functions and mechanisms of macrophages derived from PGRN gene knockout(PGRN)C57BIV6 mice in the invasion and migration of breast cancer cells.Methods Breast cancer cells were cultured in conditioned medium of macrophages derived from WT and PGRN^-/-mice.Transwell™assay and scratch assay were used to detect the invasion and migration ability of cancer cells.Western blot analysis was used to detect the expression of E-cadherin and N-cadherin in cancer cells.Cytokine array,real-time quantitative PCR and ELISA were performed to investigate the differences of cytokines secreted by macrophages derived from WT and PGRN-/-mice.Breast cancer cells were treated by the differentially expressed cytokine interleukin_6(IL-6),and then the above methods were used to investigate its effect on cancer cells.Western blot analysis was used to verify the roles of NF-kB and JAK/STAT3 signaling pathways.Results The macrophages derived from PGRN"mice blocked NF-kB signaling pathway,reduced IL-6 secretion,and inhibited the invasion and migration of breast cancer cells.IL-6 activated JAK/STAT3 signaling pathway to promote the invasion and migration of breast cancer cells.Conclusion The macrophages derived from PGRN_/_mice can block the NF-kB and JAK/STAT3 signaling pathways,down-regulate IL-6 expressi on,and in hibit the in vasi on and migration of breast can cer cells.